製品の概要

  • 製品名Anti-ROCK2 antibody
    ROCK2 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to ROCK2
  • アプリケーション適用あり: IP, ICC/IF, IHC-Fr, WB, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Human
    交差が予測される動物種: Cow, Chimpanzee, Rhesus monkey
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human ROCK2.

    (Peptide available as ab71597.)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following Lysates: Mouse Liver Tissue SK N SH Whole Cell U2OS Whole Cell HeLa Whole Cell Rat Liver Tissue This antibody gave a positive result in IHC in the following FFPE tissue: Human liver cancer. ICC/IF: HAP1-ROCK2 WT and KO cells

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab71598 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
IP Use a concentration of 5 µg/ml.
ICC/IF 1/250.
IHC-Fr Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 171 kDa (predicted molecular weight: 161 kDa).Can be blocked with Human ROCK2 peptide (ab71597).
IHC-P Use a concentration of 5 µg/ml.

ターゲット情報

  • 機能Regulates the assembly of the actin cytoskeleton. Promotes formation of stress fibers and of focal adhesion complexes. Plays a role in smooth muscle contraction.
  • 配列類似性Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family.
    Contains 1 AGC-kinase C-terminal domain.
    Contains 1 PH domain.
    Contains 1 phorbol-ester/DAG-type zinc finger.
    Contains 1 protein kinase domain.
    Contains 1 REM (Hr1) repeat.
  • 翻訳後修飾Phosphorylated upon DNA damage, probably by ATM or ATR.
  • 細胞内局在Cytoplasm. Cell membrane. Cytoplasmic, and associated with actin microfilaments and the plasma membrane.
  • Information by UniProt
  • 参照データベース
  • 別名
    • coiled-coil-containing protein kinase 2 antibody
    • KIAA0619 antibody
    • p164 ROCK 2 antibody
    • p164 ROCK-2 antibody
    • Rho associated coiled coil containing protein kinase 2 antibody
    • Rho associated protein kinase 2 antibody
    • Rho associated, coiled coil containing protein kinase II antibody
    • Rho kinase 2 antibody
    • Rho-associated antibody
    • Rho-associated protein kinase 2 antibody
    • ROCK 2 antibody
    • Rock II antibody
    • Rock2 antibody
    • ROCK2_HUMAN antibody
    • Rock2m antibody
    • ROK alpha antibody
    • ROKalpha antibody
    see all

Anti-ROCK2 antibody 画像



  • Predicted band size : 161 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: ROCK2 knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: A10 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab71598 observed at 171 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab71598 was shown to recognize ROCK2 when ROCK2 knockout samples were used, along with additional cross-reactive bands. Wild-type and ROCK2 knockout samples were subjected to SDS-PAGE. ab71598 and ab8245 (loading control to GAPDH) were diluted 1 μg/mL and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • ab71598 staining ROCK2 in wild-type HAP1 cells (top panel) and ROCK2 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab71598 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

  • All lanes : Anti-ROCK2 antibody (ab71598) at 1 µg/ml

    Lane 1 : Liver (Mouse) Tissue Lysate
    Lane 2 : SK N SH (Human neuroblastoma) Whole Cell Lysate
    Lane 3 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 5 : Liver (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 161 kDa
    Observed band size : 171 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 100 kDa,125 kDa. We are unsure as to the identity of these extra bands.
  • ROCK2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to ROCK2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab71598.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 171kDa; ROCK2
  • ICC/IF image of ab71598 stained HeLa cells. The cells were 10% neutral buffered formalin fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab71598, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • IHC image of ROCK2 staining in Human liver cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab71598, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-ROCK2 antibody (ab71598) at 1/1000 dilution

    Lane 1 : Human primary myometrial whole cell lysate with scrambled siRNA
    Lane 2 : Human primary myometrial whole cell lysate with scrambled siRNA
    Lane 3 : Human primary myometrial whole cell lysate with siRNA against ROCK2
    Lane 4 : Human primary myometrial whole cell lysate with siRNA against ROCK2

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP conjugated Goat anti-rabbit IgG polyclonal at 1/4000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 161 kDa
    Observed band size : 160-170 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes

    This image is courtesy of an anonymous Abreview

    Blocked with 5% BSA for 1 hour at 20°C

    See Abreview

Anti-ROCK2 antibody (ab71598) 使用論文

This product has been referenced in:
  • Hudson CA  et al. Phasic contractions of isolated human myometrium are associated with Rho-kinase (ROCK)-dependent phosphorylation of myosin phosphatase-targeting subunit (MYPT1). Mol Hum Reprod 18:265-79 (2012). WB . Read more (PubMed: 22155728) »

See 1 Publication for this product

Product Wall

Abreviews
Application Western blot
Loading amount 50 µg
Gel Running Conditions Non-reduced Non-Denaturing (Native) (12)
Sample Rat Tissue lysate - whole (brain)
Specification brain
Treatment Brain trauma
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Da Liu

Verified customer

投稿 Mar 18 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (primary myometrial cells)
Loading amount 20 µg
Specification primary myometrial cells
Treatment scrambled (lanes 1 and 2) or ROCK2 siRNA (lanes 3 and 4)
Gel Running Conditions Reduced Denaturing (9% Bis-Tri gel)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

投稿 May 14 2012

Thank you for your enquiry.

This antibody should bind to phophorylated ROCK2. The epitope ab71598 binds is within the first 100 amino acid of ROCK2 and the potentially phophorylated residues are located more in the C-terminal half of the prot...

Read More

Thank you for contacting us.

The immunogen of this antibody falls between 7-23 amino acid residues of human ROCK2 protein. The antibody cannot be used to distinguish phopho protein from non phospho protein. This antibody will bind to the 7-2...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (heart)
Specification heart
Fixative Acetone
Permeabilization Yes - 0.2% Triton X-100
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

投稿 Jun 17 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"